JP2004532385A - Systems and methods for in vitro analysis of therapeutic agents - Google Patents

Abstract

A system for in vitro analysis of a therapeutic agent is provided.
The system includes a first flow cell having a reservoir for holding a therapeutic agent, a first cell chamber adapted to receive at least a first sample of the therapeutic agent, A second flow cell having a second cell chamber adapted to receive at least a second sample of the therapeutic agent, wherein the first flow cell has a first passage length (b _c ^') has, above the second flow cell _"includes ^a) a first path length of said sensitivity coefficient (f) × b ^_c" second path length (b c substantially equal, The system further comprises a membrane chamber having a biological cell membrane therein for receiving at least a third sample of the therapeutic agent, and wherein the membrane potential of the biological cell membrane is detected. And the first and second Formed by and a spectrum detecting means for detecting the spectral characteristics of the therapeutic agent samples.

Description

【Technical field】
[0001]
The present invention relates generally to electrophysiological testing of therapeutic agents. In particular, the invention relates to systems and methods for in vitro testing of therapeutic agents using spectroscopic means to accurately determine the concentration of the therapeutic agent.
[Background Art]
[0002]
The potential of cardiovascular and non-cardiovascular therapeutics or agents to prolong the electrocardiographic QT interval (ie, cardiac repolarization time between two ventricular arrays) has been an important factor in the development of new therapeutics The same is true in the future. In fact, a variety of non-cardiovascular therapeutic agents, which are unexpected in terms of the mechanism of action that prolongs QT, cause a significant number of serious cardiac accidents. Such drugs belong to different pharmacological classes. For example, eutrophic agents (eg, tricyclic amitriptyline and tetracyclic antidepressants, phenothiazine derivatives, haloperidol, pimozide, risperidone, sertindole, etc.), prokinetic (eg, cisapride), antimalarial Agents (eg, halofantrine, quinine, chloroquine), antibiotics belonging to several chemical families (eg, azithromycin, erythromycin, clarithromycin, spiramycin, pentamidine, trimethoprim-sulfamethoxazole, sparfloxa) Shin), antibacterial agents (e.g., ketoconazole, Fukonazoru, itraconazole), drugs for urinary incontinence (e.g., terodiline), certain histamine H ₁ - receptors antagonists (e.g., astemizole, terfenadine, Jifenhidorami ), And the like.
[0003]
These therapies may, in some very rare cases, cause polymorphic cardiomyopathy such as torsades de pointes in the presence of ancillary factors that directly or indirectly cause arrhythmias. It causes a beat. Related factors include congenital or acquired long QT syndrome, ischemic heart disease, congestive heart disease, severe liver or kidney dysfunction, bradycardia, electrolyte parallelism (eg, low potassium due to diuretic treatment) Blood glucose, hypomagnesemia, hypocalcemia, acidosis, intercellular Ca ⁺⁺ load), intentional or accidental overdose, combination therapy with ion channel blockers that suppress the drug detoxification process, and the like.
[0004]
Accordingly, several preclinical tests have been employed to determine the cardiovascular effects of the provided therapeutic agents. Well-known techniques for determining the concentration of a provided therapeutic agent consist primarily of high performance liquid chromatography (HPLC) or other analytical tests, which, unless preconcentrated or used in large quantities, Generally limited to high therapeutic drug concentrations.
[0005]
However, it is generally known that these known physiological assays are often cumbersome, expensive, time consuming and frustrated by technical problems. In addition, the above-described HPLC and / or analytical tests require hours or days to analyze the sample and process the data, depending on the complexity and number of samples.
[0006]
Accordingly, one object of the present invention is to provide a system and method for in vitro analysis of therapeutic agents that is fast and economical.
[0007]
It is another object of the present invention to provide systems and methods for in vitro analysis of low concentrations of therapeutic agents.
[0008]
It is yet another object of the present invention to provide a system and method for correlating the electrophysiological effects of a provided therapeutic agent over a wide range of concentrations.
[0009]
(Summary of the Invention)
In accordance with the above objects and preferred embodiments set forth below, a system for in vitro analysis of a therapeutic agent is provided by the present invention. That is, the system includes a first flow cell having a reservoir for holding a therapeutic agent, a first cell chamber adapted to receive at least a first sample of the therapeutic agent, and at least one of the therapeutic agents. A second flow cell having a second cell chamber adapted to receive a second sample, wherein the first flow cell has a first passage length (b _c ^′ ). , The second flow cell has a second path length (b _c ^″ ), the first path length is substantially equal to a sensitivity factor (f) × b _c ^″ , and the system further comprises the treatment A membrane chamber having a biological cell membrane therein for receiving at least a third sample of a drug, wherein the membrane chamber is adapted to detect a membrane potential of the biological cell membrane; Examine the spectral properties of a therapeutic drug sample ; And a spectrum detecting means for comprising.
[0010]
A method for in vitro analysis of a therapeutic agent according to the present invention comprises: (i) introducing a first sample of a therapeutic agent into a first flow cell having a first path length (b _c ^′ ); (Ii) introducing a second sample of the therapeutic agent into a second flow cell having a second path length (b _c ^″ ), wherein the first path length (b _c ^′ ) is (Iii) introducing a third sample of the therapeutic agent into a membrane chamber having a biological cell membrane therein, and (iv) introducing a third sample of the therapeutic agent substantially equal to the sensitivity factor (f) × b _c ^″ . Measuring an absorption spectrum of the first therapeutic agent sample by transmitting a given wavelength of light to the first flow cell; and (v) converting a given wavelength of second light to the first flow cell. Measuring the absorption spectrum of the second therapeutic agent sample by transmitting it to a second flow cell; Comprising comprising a step of detecting the membrane potential of vi) biological cell membranes.
BEST MODE FOR CARRYING OUT THE INVENTION
[0011]
The methods and systems of the present invention substantially reduce or eliminate the shortcomings and deficiencies associated with conventional electrophysiological testing of therapeutic agents. As will be described in more detail below, the system generally comprises introducing a plurality of flow cells, a spectroscopic analysis means communicating with the flow cells, a membrane chamber means, and a related therapeutic agent into the flow cells and the membrane chamber means. And a flow path means for performing the operation.
[0012]
Here, the “therapeutic agent” is a broad concept including active ingredients or ingredients (ingredients, components, elements) of a pharmaceutical composition, a drug, a pharmaceutical and the like.
[0013]
Cardiac action potential (or membrane potential) is generally defined as a pattern of electrical activity associated with excitable biological cells (eg, heart cells). It is generated by differentiated membrane structures, such as ion pumps, ion exchangers, and most importantly, biologically important ions (Na ⁺ , Ca ⁺⁺ and K ⁺ ) passing through voltage-gated ion channels. FIG. 4 illustrates the result of a number of individual, continuously activated currents. It is thought that such a current depolarizes when an extracellular positive charge is carried into a cell, and repolarizes when a positive charge is carried outside the cell.
[0014]
Therapeutic agents that alter the normal flow of ions through the channel have the potential to alter, or often alter, certain aspects of the membrane potential, and thus affect cardiac function. In fact, Na ⁺ channel blockers reduce the rate of rise of membrane potential (V max), disrupting cardiac performance and, if severe, can be life threatening. Agents that reduce the rate of inactivation of Na currents and increase residual Na currents prolong the duration of the action potential (ADP), prolong the QT interval, and thus reduce torsades de pointes arrhythmias. May cause it to occur. Ca ⁺⁺ channel blockers generally reduce ADP, decrease AV conduction velocity, and cause cardiac decline. In contrast, Ca ⁺⁺ channel activators prolong ADP and can cause arrhythmias. Finally, K ⁺ channel blockers prolong ADP and QT and can cause arrhythmias, and K ⁺ channel activators shorten ADP and can cause arrhythmias.
[0015]
Thus, an effective indicator (or parameter) of a provided therapeutic agent for adverse cardiac effects is a change in membrane potential (ie, membrane resting potential) resulting from the introduction or exposure to the therapeutic agent. . In fact, this parameter has often been subject to conventional physiologic testing of the offered therapeutic (or potential drug). As described in detail below, this parameter is also used in the present invention for assessing the potential physiological effects of a therapeutic agent.
[0016]
Referring to FIG. 1, there is schematically shown an example of an in vitro analysis system 5 of the present invention. As shown in FIG. 1, the system 5 preferably includes a storage means (eg, a storage unit) 10, an optical spectroscopic detection means 20, and a plurality of flow cells (preferably first and second flow cells 60, 70). , A membrane chamber means 40 and a flow path means 12.
[0017]
According to the invention, the storage means 10 is designed to be able to store at least one diluted therapeutic agent. As will be apparent to those skilled in the art, the storage means 10 can be of various capacities (and shapes) within the scope of the present invention. In a preferred embodiment of the invention, the volume of this storage means 10 is selected in the range of 1 to 2000 ml.
[0018]
This storage means 10 is designed to receive the flow path means 12 of the present invention. The channel means 12 preferably comprises a substantially non-adsorbent tube (eg, stainless steel, PEEK®). As shown in FIG. 1, a pump means 14 is also provided to facilitate the flow of the therapeutic agent (ie, the therapeutic agent sample) from the storage means 10 to the first and second flow cells 60, 70. . The pump means 14 is in communication with the flow path means 12 and is preferably arranged between the storage means 10 and the flow cells 60,70. According to the present invention, the pump means 14 is used to provide a sample flow rate in the range of 0.5 to 10 mL / min, more preferably in the operating range of about 2 to 5 mL / min.
[0019]
As will be apparent to those skilled in the art, it is possible to provide the above-described sample flow rate using various forms of the pump means 14 within the scope of the present invention. In a preferred example, this pump means 14 comprises a peristaltic pump.
[0020]
Referring to FIG. 2, an example of a first flow cell 60 of the present invention is shown. For simplicity, only the first flow cell 60 is shown here. However, it should be understood that the second flow cell 70 of the above embodiment may be constructed in a similar manner, and that the description of the first flow cell 60 applies equally to each of the flow cells 60, 70. .
[0021]
As shown in FIG. 2, the flow cell 60 preferably comprises a substantially cylindrical body 62, which is provided with an inlet port 64, an outlet port 66, and a cell chamber indicated generally at 63, and Samples can be received. According to the present invention, the flow path 61 is defined by the inlet port 64, the outlet port 66, and the cell chamber 63.
[0022]
As will be apparent to those skilled in the art, various flow cell bodies 62 can be used within the scope of the present invention. In a preferred embodiment of the present invention, the main body 62 of each of the flow cells 60 and 70 includes a core portion made of a material such as a polymer, silica, or a chalcogenide, and a clad 67 provided on an outer surface of the core portion. The clad 67 is made of a material such as a polymer and doped silica (see FIGS. 2 and 4).
[0023]
According to the present invention, the first flow cell 60 is the first optical transmission means 21a, which supplies excitation light or radiation of a predetermined wavelength (and / or a predetermined range thereof) to the first flow cell chamber. 63 (ie, a first sample) and a first light detecting means 23a for detecting light transmitted from the first sample. Similarly, the second flow cell 70 is also the second optical transmission unit 21b, and supplies the excitation light having a predetermined wavelength (and / or a predetermined range thereof) to the sample in the cell chamber of the second flow cell 70. (Ie, a second sample) and second light detecting means 23b for detecting light transmitted from the second sample.
[0024]
Referring to FIG. 3, another example of the flow cell 80 of the present invention is shown. The flow cell 80 is similarly provided with an inlet port 82, an outlet port 84 and a cell chamber indicated generally at 86 therebetween for receiving a therapeutic agent sample.
[0025]
The first flow cell 80 is a first optical transmission means 85 for providing excitation light or radiation of a predetermined wavelength to the flowable therapeutic agent in the flow cell chamber 86 (ie, the first sample). And first light detecting means 87 for detecting light transmitted from the first sample.
[0026]
It is known that as each sample passes through a respective flow cell chamber (eg, 63), the amount of excitation light transmitted through the flow cell chamber 63 decreases according to Beer's law. That is,
Equation 1 A = I / I ₀ = 10− ^∝bc
here,
A = absorbance I = transmitted radiation force I ₀ = incident radiation force ∝ = molar extinction coefficient of the sample c = sample concentration (mol / liter)
b = path length of light in the chamber (cm)
The absorbance (A) is thus defined as the product of ∝bc. According to Beer's law, the absorbance (A) is further proportional to both the sample concentration (c) and the light path length (b).
[0027]
As will be apparent to those skilled in the art, the optical path length (b) is generally considered to be a "linear path length", which is applicable to an optical transmission / detection configuration as shown in FIG. Further, as will be apparent to those skilled in the art, the above Beer's law also applies to the optical transmission / detection configuration (ie, 21a, 23a) of the tubular flow cell 60 (see FIG. 2) of the present invention employing attenuated total reflection. It is possible.
[0028]
Referring to FIG. 4, if the radiation (light), indicated generally at 100, has total internal reflection 102, it is known that some of the wavelengths actually penetrate beyond the reflective surface and into the medium (or sample). ing. The penetration depth is generally expressed in d _p.
[0029]
Since the penetration depth d _p is defined as unit length / reflection, the equivalent path length (b _c ) can be obtained as follows.
[0030]
Formula _{2 b c = d p × R} × L
here,
d _p = penetration depth per reflection R = number of reflections per unit length L = tube or cell length Equivalent path length (b _c ) by applying equation 1 to the absorbance of each sample (A) Can be obtained.
[0031]
As will be apparent to those skilled in the art, the effective path length (b _c ) of each of the flow cells 60, 70 depends directly on the length of the flow cell body 62. That is, different path lengths are provided by adjusting the lengths of the flow cells 60 and 70, respectively.
[0032]
From Beer's law, if the absorbance range of a spectrophotometer (eg, a spectrophotometer) is constant (this is a common component of conventional spectrophotometers), because of the lower molar extinction coefficient (∝), or It can be inferred that the path length (b or b _c ) must be increased for a lower concentration (c). However, as is known to those skilled in the art, the response band generally associated with longer path lengths is too narrow and undesirable, and will therefore be limited.
[0033]
Furthermore, it is well known that a wider response band can be achieved by employing two flow cells with different path lengths (b _c ^′ , b _c ^″ ). 1 to 100 cm.
[0034]
However, the inventors have found that if b _c ^′ is substantially equal to (b _c ^″ × sensitivity factor (f)), the best dynamic (ie, substantially linear) response range can be achieved. According to the invention, the sensitivity factor (f) is preferably in the range from 1 to 100, more preferably in the range from 1 to 20.
[0035]
Referring to FIG. 5, there is shown an absorbance spectrum for a low concentration of therapeutic agent (ie, 〜18 ng / mL) and an absorbance spectrum for a high concentration of therapeutic agent (ie, ３3800 ng / mL). These absorbance spectra were obtained using a first flow cell having a path length of about 5 cm and a second flow cell having a path length of about 50 cm (ie, f = 10).
[0036]
Referring to curve A, it can be seen that a path length of 5 cm is insufficient to detect low concentrations of therapeutic agent over the wavelength range of 200 nm to 400 nm. However, as shown in curve B, the same low concentration of therapeutic agent can be easily detected (and more quantifiable) at a path length of 50 cm.
[0037]
Referring to curve C, it will be seen that high concentrations of therapeutic agent cannot be quantified at a path length of 50 cm. However, as shown by curve D, the same high concentration of therapeutic agent can be easily detected with a path length of 5 cm.
[0038]
Thus, in a preferred embodiment of the present invention, the first flow cell 60 has an effective path length (b _c ^′ ) in the range of 0.1 to 100 cm, and the second flow cell 70 has an effective path length in the range of 0.1 to 100 cm. more preferably has road length _(b ^{c ").,} the first flow cell 60 has an effective path length in the range of about 5 to 50cm _{(b c} ^'), the second flow cell 70 about 50 It has an effective path length (b _c ^″ ) in the range of -100 to 100 cm. We have found that the exact spectral properties of therapeutic agents having substantially low concentrations in the range of 5 to 10 ng / mL can be readily detected by path lengths ( _bc ^' , _bc ^" ) in the above ranges. Was done.
[0039]
Referring to FIG. 6, in addition to the first and second light transmitting means 21a and 21b and the first and second light detecting means 23a and 23b shown in FIG. 2, the spectral detecting means 20 of the present invention is further preferable. A light source means 22 for providing light of a wavelength to the first and second light transmission means 21a and 21b via optical lines 24a and 24b, and a light detected by the first and second light detection means 23a and 23b. Analyzer means 26 for analyzing. The first and second light detecting means 23a and 23b communicate with the analyzer means 26 via optical lines 28a and 28b, respectively.
[0040]
As yet another possible embodiment of the invention, the storage means 10 further comprises a light transmission means 25a and a light detection means 25b, which are connected via optical lines 27a, 27b to the spectroscopic analysis means 20 of the invention. Operably connected to As will be apparent to those skilled in the art, the light transmitting means 25a and the light detecting means 25b provide a means for simultaneously examining and monitoring the therapeutic agent in the storage means 10, thus detecting loss of the therapeutic agent, It provides a means to ensure that the sample analyzed in the flow cells 60, 70 is representative of the therapeutic agent contained in the storage means 10.
[0041]
As shown in FIG. 6, the spectroscopic analysis means 20 preferably includes the detected spectroscopic analysis characteristics of the first and second samples (and the raw material chemicals contained in the storage means 10) and the spectroscopic analysis means of the present invention. Means 30 for storing at least control parameters for the sample, the processor means 32 for processing at least the spectroscopic properties of the sample (and the raw chemical), and the sample and raw chemical contained in the storage means 10. A first display means 34 (indicated by phantom lines) for displaying at least the spectral properties as well as displaying the "processed" spectral properties (e.g., the average value) of the sample.
[0042]
As will be apparent to those skilled in the art, a variety of conventional light source means 22 and / or analysis means 26 may be used within the scope of the present invention, thereby providing a wavelength range of light and providing first and second light sources. The spectral characteristics (for example, the absorption spectrum of the absorbed light) obtained by the detection units 23a and 23b may be analyzed to control the spectral analysis unit 20 of the present invention. The analysis means in this case is described in US Pat. 4,664,522 and MCS-521 Fiber Optics UV / VIS Spectrophotometer (Carl Zeiss), which is incorporated herein by reference. The analyzing means 26 and / or the processor means 32 may comprise a personal computer.
[0043]
Referring to FIG. 7, there is shown a membrane chamber means 40 of the present invention. The membrane chamber means 40 has a membrane chamber main body 42 into which an infusion inlet 44, an infusion outlet 46, a dispensing well 48, a membrane well 50, and a diffusion plate 52 disposed between the dispensing well 48 and the membrane well 50. Is provided.
[0044]
As shown in FIG. 7, the membrane chamber means 40 further has a membrane means 52 arranged in the membrane well 50. As used herein, the term "membrane means" refers to a biological cell membrane, including sheets or layers of biological organs, ventricular and / or papillary muscle and Purkinje fibers.
[0045]
In a preferred embodiment of the present invention, the membrane means 52 comprises Purkinje fibers. As is known, Purkinje fibers are often used in electrobiological tests because the primary ionic current underlying their action potential contributes to the repolarization process of the human heart Because it is similar to
[0046]
In order to check the membrane potential of the membrane means 52, the membrane chamber means 40 further has a plurality of electrodes. Referring to FIG. 7, at least one, and preferably two, bipolar electrodes 54a, 54b are operably connected to a membrane means 52 near one end thereof. These electrodes 54a, 54b are further connected to the stimulating means 55 of the present invention via the leads 54a, 54b (see FIG. 1) so as to apply a stimulating charge or current to the membrane means 52.
[0047]
As shown in FIG. 7, another electrode is further provided. As a preferred example, this electrode is an intercellular microelectrode 56, which is also operably connected to the membrane means 52.
[0048]
According to the invention, this microelectrode 56 is designed to detect the membrane potential of the membrane means 52. Preferably, this microelectrode 56 communicates (via a lead 57c) with a second display means 58 that visually indicates or indicates the detected membrane potential (see FIG. 1).
[0049]
Examination of the therapeutic agent according to the invention is preferably carried out as follows. The spectral analysis means of the present invention is first calibrated by conventional means. The means include analysis of a blank (or primary) sample as a UV reference, and analysis of a calibration sample using the respective blank sample as a reference.
[0050]
After the calibration of the spectroscopic analysis means 20, the flow (preferably diluted) of the therapeutic agent is started, and the therapeutic agent is introduced into and passed through the channel means 12 via the pump means 14. The therapeutic agent is then introduced and passed through the first and second flow cells 60, 70 and the channel 61 of the membrane chamber means 40. Note that the first and second flow cells 60 and 70 are preferably connected in series.
[0051]
The spectral characteristics of the therapeutic agent (that is, the first sample) present in the first flow cell 60 and the therapeutic agent (that is, the second sample) that exists in the second flow cell 70 correspond to the spectral analysis means 20 of the present invention. (Preferably substantially simultaneously). This spectral characteristic is then processed and analyzed by conventional means.
[0052]
In a preferred example, substantially simultaneously with the spectroscopic analysis, while the therapeutic agent is in the membrane well 50 of the membrane chamber means 40 (ie, the third sample), the membrane means 52 activates the stimulating electrodes 54a, 54b. Exposed to the initial current. Next, the membrane potential is detected via the electrode 56, and this is displayed on the second display means 58 of the present invention. The change in membrane potential is then easily determined by comparing the detected membrane potential with the potential of membrane means 52 prior to exposure to the therapeutic agent.
[0053]
As will be apparent to those skilled in the art, the systems and methods of the present invention described above may be applied to the analysis of pharmaceutical compositions containing therapeutic agents, particularly those containing substantially lower concentrations of therapeutic agents. it can.
[0054]
Overview:
From the above description, those skilled in the art can easily confirm that the present invention has the following advantages.
[0055]
1. Fast, accurate and economical (ie, low cost) in vitro analysis of therapeutics.
[0056]
2. Fast, efficient in vitro analysis of therapeutics in pharmaceutical compositions that include substantially low concentrations (ie, 5-10 ng / mL) of the therapeutic.
[0057]
3. A fast, efficient means for correlating the electrophysiological effects of therapeutic agents provided over a wide range of concentrations.
[0058]
4. A means for testing and monitoring the stability of a therapeutic agent during in vitro analysis.
[0059]
5. Means for testing and monitoring drug loss (eg, glassware absorption and / or adhesion) during in vitro analysis.
[0060]
6. Means for quickly and efficiently detecting carryover (ie, cross-contamination) in storage and / or supply lines.
[0061]
It will be apparent that those skilled in the art can make various modifications, variations and adaptations to the various uses and conditions without departing from the spirit and scope of the invention. It is therefore understood that such changes and modifications should be properly and fairly included in the equivalent scope of the appended claims.
[Brief description of the drawings]
[0062]
FIG. 1 is a schematic diagram showing an example of an in vitro spectroscopic analysis system for analyzing a therapeutic agent according to the present invention.
FIG. 2 is a partially cutaway plan view showing one example of a flow cell according to the present invention.
FIG. 3 is a partially cutaway plan view showing another example of the flow cell according to the present invention.
FIG. 4 is a schematic perspective view of a flow cell main body showing an attenuation optical path according to the present invention.
FIG. 5 is a graph showing the absorbance spectrum of a pharmaceutical composition having a low concentration therapeutic agent and a high concentration therapeutic agent, respectively.
FIG. 6 is a schematic diagram showing the spectroscopic analysis means of the present invention.
FIG. 7 is a partially cutaway perspective view showing a membrane chamber means of the present invention.

Claims (23)

A reservoir for holding a therapeutic agent;
A first flow cell in communication with the reservoir having a first cell chamber adapted to receive at least a first sample of the therapeutic agent;
A second flow cell in communication with the reservoir having a second cell chamber adapted to receive at least a second sample of the therapeutic agent;
Here, the first flow cell has a first passage length (b _c ^′ ), the second flow cell has a second passage length (b _c ^″ ), and the first passage length is Substantially equal to the sensitivity factor (f) × b _c ^″ ;
A membrane chamber in communication with a reservoir having a biological cell membrane therein, the membrane chamber receiving at least a third sample of the therapeutic agent and detecting a membrane potential of the biological cell membrane. When;
Spectrum detecting means for detecting spectral characteristics of the first and second therapeutic agent samples;
A system for in vitro analysis of a therapeutic agent comprising: A first light transmitting means for transmitting a first light of a given wavelength into the first cell chamber; and a first transmission from the first therapeutic agent sample. First light detecting means for detecting light, second light transmitting means for transmitting a second light of a given wavelength into the second cell chamber, and the second treatment The system of claim 1, further comprising second light detection means for detecting a second transmitted light from the drug sample. Communicating with the first and second light transmitting means and the first and second light detecting means for providing the first and second light and analyzing the transmitted first and second light; 3. The system according to claim 2, further comprising control means for performing the operation. The system of claim 1, wherein said sensitivity factor has a value in the range of 1 to 100. The system according to claim 4, wherein said sensitivity factor has a value in the range of 1 to 20. The system of claim 1, wherein said first and second path lengths are in the range of 0.1 to 100 cm. 7. The system of claim 6, wherein said first path length is in the range of 5 to 50 cm. 7. The system of claim 6, wherein said second path length is in the range of 50 to 100 cm. A third light transmitting means for transmitting a third light of a given wavelength into the storage, and a third transmission light from a therapeutic agent in the storage; 3. The system of claim 1, further comprising third light detection means for communicating with said control means, said third light transmission means and said third light detection means communicating with said control means. 2. The system of claim 1, wherein said spectral detection means comprises first display means for displaying at least spectral properties of said first and second therapeutic agent samples. 2. The system of claim 1, wherein said membrane chamber has second display means for displaying a membrane potential of said biological cell membrane. A reservoir for receiving a therapeutic agent;
A first flow cell communicating with the reservoir having a first cell chamber adapted to receive at least a first sample of the therapeutic agent, and having an effective passage length in a range of about 5 to 50 cm;
A second flow cell communicating with the reservoir having a second cell chamber adapted to receive at least a second sample of the therapeutic agent and having an effective passage length in the range of about 50 to 100 cm;
A membrane chamber in communication with a reservoir having a biological cell membrane therein, the membrane chamber receiving at least a third sample of the therapeutic agent and detecting a membrane potential of the biological cell membrane. When;
Spectrum detecting means for detecting spectral characteristics of the first and second therapeutic agent samples;
A system for in vitro analysis of a therapeutic agent comprising: A first light transmitting means for transmitting a first light of a given wavelength into the first cell chamber; and a first transmission from the first therapeutic agent sample. First light detecting means for detecting light, second light transmitting means for transmitting a second light of a given wavelength into the second cell chamber, and the second treatment 13. The system of claim 12, further comprising second light detection means for detecting a second transmitted light from the drug sample. Communicating with the first and second light transmitting means and the first and second light detecting means for providing the first and second light and analyzing the transmitted first and second light; 14. The system according to claim 13, further comprising control means for performing the operation. A third light transmitting means for transmitting a third light of a given wavelength into the storage, and a third transmission light from the therapeutic agent in the storage; 15. The system of claim 14, further comprising third light detection means for communicating with said control means, said third light transmission means and said third light detection means being in communication with said control means. 16. The system of claim 15, wherein said spectral detection means comprises display means for displaying at least spectral properties of the therapeutic agent in the reservoir and the first and second therapeutic agent samples. A method for in vitro analysis of a therapeutic agent, comprising:
Introducing at least a first sample of the therapeutic agent into a first flow cell having a first path length (b _c ^′ );
Introducing at least a second sample of the therapeutic agent into a second flow cell having a second path length (b _c ^″ );
Wherein the first path length is substantially equal to the sensitivity factor (f) × b _c ^″ ;
Introducing at least a third sample of the therapeutic agent into a membrane chamber having a biological cell membrane therein;
Measuring an absorption spectrum of the first therapeutic agent sample by transmitting a given wavelength of first light to the first flow cell;
Measuring an absorption spectrum of the second therapeutic agent sample by transmitting a given wavelength of second light to the second flow cell;
Detecting the membrane potential of the biological cell membrane;
A method comprising: 18. The method of claim 17, wherein the absorption spectrum of the first therapeutic agent sample and the absorption spectrum of the second therapeutic agent sample are measured substantially simultaneously. 18. The method of claim 17, wherein said sensitivity factor has a value in the range of 1 to 100. 18. The method according to claim 17, wherein said sensitivity factor has a value in the range of 1 to 20. 18. The method of claim 17, wherein said first and second path lengths are in the range of 0.1 to 100 cm. 22. The method of claim 21, wherein said first path length is in the range of 5 to 50 cm. 22. The method of claim 21, wherein said second path length is in the range of 50 to 100 cm.

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